LOCUS       Exported               12959 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pDOE-08, complete sequence.
ACCESSION   KM507049
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12959)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Significant reduction of BiFC non-specific assembly facilitates in 
            planta assessment of heterotrimeric G-protein interactors
  JOURNAL   Plant J. (2014) In press
  PUBMED    25187041
REFERENCE   2  (bases 1 to 12959)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-2014) Department of Biology, The Pennsylvania 
            State University, 208 Mueller Laboratory, University Park, PA 16802,
            USA
REFERENCE   3  (bases 1 to 12959)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12959
                     /organism="Binary vector pDOE-08"
                     /mol_type="other DNA"
                     /clone="p4489"
                     /db_xref="taxon:1544598"
     repeat_region   1..27
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    99..104
                     /label=non-unique SacI
                     /note="non-unique SacI"
     terminator      111..363
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     regulatory      complement(111..362)
                     /regulatory_class="terminator"
                     /note="MAS mannopine synthase terminator"
     misc_feature    complement(373..406)
                     /label=BAR gene remnant
                     /note="BAR gene remnant"
     misc_feature    407..412
                     /label=MCS2 KpnI site
                     /note="MCS2 KpnI site"
     CDS             complement(413..1234)
                     /codon_start=1
                     /gene="XT-Golgi-mTq2"
                     /product="XT-Golgi-mTq2"
                     /label=XT-Golgi-mTq2
                     /note="normalized for Arabidopsis"
                     /protein_id="AIN46632.1"
                     /translation="MSKRNPKILKIFLYMLLLNSLFLIIYFVFHSSSFSVSKGEELFTG
                     VVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLSWGVQ
                     CFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGI
                     DFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTP
                     IGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            complement(413..1234)
                     /gene="XT-Golgi-mTq2"
                     /label=XT-Golgi-mTq2
     misc_feature    complement(413..1129)
                     /gene="XT-Golgi-mTq2"
                     /note="similar to protein sequence for mTurquoise2 crystal 
                     structure PDB 3ZTF; characterized in PubMed ID 22434194; 
                     mTq2, mTurquoise2 specific sequence"
     misc_difference complement(692..694)
                     /gene="XT-Golgi-mTq2"
                     /label=differs from mTurquoise, mTq2=I146F
                     /note="differs from mTurquoise, mTq2=I146F"
     sig_peptide     complement(1130..1234)
                     /gene="XT-Golgi-mTq2"
                     /note="exclusive localization to the medial Golgi 
                     cisternae; from Arabidopsis AtXylT/XYLT 
                     (beta-1,2-xylosyltransferase, At5g55500); characterized in 
                     PubMed ID 12943552; confirmed by Saint-Jore-Dupas et al. 
                     (Plant Cell 2006;18;3182-3200)"
     misc_feature    1235..1240
                     /label=MCS2 NruI site
                     /note="MCS2 NruI site"
     regulatory      complement(1238..1618)
                     /regulatory_class="promoter"
                     /note="mas1-2 mannopine synthase promoter; mas2 drives 
                     MCS2 ORF; mas1 truncated by 96 nt; enhancer elements, TATA
                     box, and start codon missing"
     promoter        complement(1238..1618)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     regulatory      1631..2960
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter"
     promoter        2614..2959
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2950..2974
                     /label=pDOE mcs1F with XhoI
                     /note="pDOE mcs1F with XhoI"
     misc_feature    2962..2978
                     /label=XhoSac linker
                     /note="XhoSac linker"
     misc_feature    2979..3044
                     /label=TMV omega prime translation leader
                     /note="TMV omega prime translation leader"
     misc_feature    2989..3043
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             3045..3728
                     /codon_start=1
                     /gene="NmVen210-X"
                     /product="NmVenus210-mcs1 fusion protein"
                     /label=NmVen210-X
                     /protein_id="AIN46633.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKGASGGGSGSMGR
                     ALGTS"
     gene            3045..3728
                     /gene="NmVen210-X"
                     /label=NmVen210-X
     misc_feature    3045..3674
                     /gene="NmVen210-X"
                     /label=NmVenus210 specific sequence
                     /note="NmVenus210 specific sequence"
     CDS             3045..3563
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     misc_difference 3663..3665
                     /gene="NmVen210-X"
                     /label=A206K monomerizing mutation
                     /note="A206K monomerizing mutation"
     misc_feature    3675..3728
                     /label=MCS1 front end
                     /note="MCS1 front end"
     misc_feature    3729..3750
                     /label=MCS1 back end
                     /note="MCS1 back end"
     regulatory      3751..4476
                     /regulatory_class="terminator"
                     /note="OCS3octapine synthase terminator"
     terminator      3761..4468
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3761..3769
                     /note="similar to native ocs region; c is 14nt downstream 
                     of Agro OCS stop codon (MacDonald 1991)"
     misc_feature    4477..4482
                     /label=non-unique NdeI
                     /note="non-unique NdeI"
     regulatory      4483..5816
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter"
     promoter        5469..5814
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5810..5834
                     /label=pDOE mcs3bF with Bsu36I
                     /note="pDOE mcs3bF with Bsu36I"
     misc_feature    5817..5832
                     /label=Bsu361 linker
                     /note="Bsu361 linker"
     regulatory      5833..5898
                     /regulatory_class="enhancer"
                     /note="TMV-Omega enhancer"
     misc_feature    5843..5897
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             5899..6138
                     /codon_start=1
                     /gene="CVen210-SFS-X"
                     /product="CVenus210-mcs3 fusion protein"
                     /label=CVen210-SFS-X
                     /note="StrepII-FLAG-StrepII epitopes are spatially 
                     separated"
                     /protein_id="AIN46634.1"
                     /translation="MDPNEKRDHMVLLEFVTAAGITLGMDELYKGSGSWSHPQFEKSGG
                     DYKDDDDKASWSHPQFEKSGASGMGSLRSHVTSG"
     gene            5899..6138
                     /gene="CVen210-SFS-X"
                     /label=CVen210-SFS-X
     misc_feature    5902..5988
                     /gene="CVen210-SFS-X"
                     /label=CVenus210 specific sequence
                     /note="CVenus210 specific sequence"
     misc_feature    6001..6024
                     /gene="CVen210-SFS-X"
                     /label=Strep II tag (A)
                     /note="Strep II tag (A)"
     CDS             6001..6024
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    6034..6057
                     /gene="CVen210-SFS-X"
                     /label=FLAG tag
                     /note="FLAG tag"
     CDS             6034..6057
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    6064..6087
                     /gene="CVen210-SFS-X"
                     /label=Strep II tag (B)
                     /note="Strep II tag (B)"
     CDS             6064..6087
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    6103..6138
                     /gene="CVen210-SFS-X"
                     /label=MCS3 front end
                     /note="MCS3 front end"
     misc_feature    6139..6156
                     /label=MCS3 back end
                     /note="MCS3 back end"
     regulatory      6158..6410
                     /regulatory_class="terminator"
                     /note="NOS nopaline synthase terminator"
     terminator      6158..6410
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6420..6436
                     /label=remnant from pFGC5941
                     /note="remnant from pFGC5941"
     misc_feature    6437..12959
                     /label=derived from pCAMBIA1300 via pFGC5941
                     /note="derived from pCAMBIA1300 via pFGC5941"
     primer_bind     complement(6461..6477)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6679..6702
                     /label=right border from C58 T-DNA
                     /note="right border from C58 T-DNA"
     misc_feature    6680..6704
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      7765..8765
                     /label=pVS1
                     /note="pVS1"
     CDS             8005..8634
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9063..10136
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      10202..10396
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    10740..10880
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(11066..11654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      11169..11450
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(11741..12535)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aadA
                     /note="AphA-3"
                     /protein_id="AIN46635.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(11741..12535)
                     /gene="aadA"
                     /label=aadA